Vascular wall-resident stem cells (VW-SCs) play a critical role in maintaining normal vascular function and regulating vascular repair. Understanding the basic functional characteristics of the VW-SCs will facilitate the study of their regulation and potential therapeutic applications. The aim of this study was to establish a stable method for the isolation, culture, and validation of the CD34⁺ VW-SCs from mice, and to provide abundant and reliable cell sources for further study of the mechanisms involved in proliferation, migration and differentiation of the VW-SCs under various physiological and pathological conditions. The vascular wall cells of mouse aortic adventitia and mesenteric artery were obtained by the method of tissue block attachment and purified by magnetic microbead sorting and flow cytometry to obtain the CD34⁺ VW-SCs. Cell immunofluorescence staining was performed to detect the stem cell markers (CD34, Flk-1, c-kit, Sca-1), smooth muscle markers (SM22, SM MHC), endothelial marker (CD31), and intranuclear division proliferation-related protein (Ki-67). To verify the multipotency of the isolated CD34⁺ VW-SCs, endothelial differentiation medium EBM-2 and fibroblast differentiation medium FM-2 were used. After culture for 7 days and 3 days respectively, endothelial cell markers and fibroblast markers of the differentiated cells were evaluated by immunofluorescence staining and q-PCR. Furthermore, the intracellular Ca²⁺ release and extracellular Ca²⁺ entry signaling were evaluated by TILLvisION system in Fura-2/AM loaded cells. The results showed that: (1) High purity (more than 90%) CD34⁺ VW-SCs from aortic adventitia and mesenteric artery of mice were harvested by means of tissue block attachment method and magnetic microbead sorting; (2) CD34⁺ VW-SCs were able to differentiate into endothelial cells and fibroblasts in vitro; (3) Caffeine and ATP significantly activated intracellular Ca²⁺ release from endoplasmic reticulum of CD34⁺ VW-SCs. Store-operated Ca²⁺ entry (SOCE) was activated by using thapsigargin (TG) applied in Ca²⁺-free/Ca²⁺ reintroduction protocol. This study successfully established a stable and efficient method for isolation, culture and validation of the CD34⁺ VW-SCs from mice, which provides an ideal VW-SCs sources for the further study of cardiovascular diseases.
Mice ; Animals ; Endothelial Cells ; Cell Differentiation/physiology* ; Stem Cells ; Adventitia ; Fibroblasts ; Cells, Cultured ; Antigens, CD34/metabolism*

Humans ; Prevalence ; Mitral Valve ; Mitral Valve Prolapse ; Adventitia

Humans ; Prevalence ; Mitral Valve ; Mitral Valve Prolapse ; Adventitia

OBJECTIVES: In this study, we explored how adiponectin mediated urotensin II (UII)‍-induced tumor necrosis factor-‍α (TNF-‍α) and α‍-smooth muscle actin (α‍-SMA) expression and ensuing intracellular signaling pathways in adventitial fibroblasts (AFs). METHODS: Growth-arrested AFs and rat tunica adventitia of vessels were incubated with UII and inhibitors of signal transduction pathways for 1‍‒‍24 h. The cells were then harvested for TNF-α receptor (TNF-‍α-R) messenger RNA (mRNA) and TNF-‍α protein expression determination by reverse transcription-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA), respectively. Adiponectin and adiponectin receptor (adipoR) expression was measured by RT-PCR, quantitative real-time PCR (qPCR), immunohistochemical analysis, and cell counting kit-8 (CCK-8) cell proliferation experiments. We then quantified TNF-α and α-SMA mRNA and protein expression levels by qPCR and immunofluorescence (IF) staining. RNA interference (RNAi) was used to explore the function of the adipoR genes. To investigate the signaling pathway, we applied western blotting (WB) to examine phosphorylation of adenosine 5'-monophosphate (AMP)‍-activated protein kinase (AMPK). In vivo, an adiponectin (APN)‍-knockout (APN-KO) mouse model mimicking adventitial inflammation was generated to measure TNF-α and α‍-SMA expression by application of qPCR and IF, with the goal of gaining a comprehensive atlas of adiponectin in vascular remodeling. RESULTS: In both cells and tissues, UII promoted TNF-α protein and TNF-α-R secretion in a dose- and time-dependent manner via Rho/protein kinase C (PKC) pathway. We detected marked expression of adipoR1, T-cadherin, and calreticulin as well as a moderate presence of adipoR2 in AFs, while no adiponectin was observed. Globular adiponectin (gAd) fostered the growth of AFs, and acted in concert with UII to induce α-SMA and TNF-α through the adipoR1/T-cadherin/calreticulin/AMPK pathway. In AFs, gAd and UII synergistically induced AMPK phosphorylation. In the adventitial inflammation model, APN deficiency up-regulated the expression of α-SMA, UII receptor (UT), and UII while inhibiting TNF-‍α expression. CONCLUSIONS From the results of our study, we can speculate that UII induces TNF‍-‍α protein and TNF-‍α‍-R secretion in AFs and rat tunica adventitia of vessels via the Rho and PKC signal transduction pathways. Thus, it is plausible that adiponectin is a major player in adventitial progression and could serve as a novel therapeutic target for cardiovascular disease administration.
Mice ; Rats ; Animals ; Adventitia/metabolism* ; Tumor Necrosis Factor-alpha/metabolism* ; Calreticulin/metabolism* ; Vascular Remodeling ; AMP-Activated Protein Kinases/metabolism* ; Cells, Cultured ; RNA, Messenger/genetics* ; Inflammation

Adventitia ; Atherosclerosis/genetics* ; Cell Communication ; Humans ; Hyperhomocysteinemia/genetics* ; Sequence Analysis, RNA

Adventitia ; Humans ; Mitral Valve ; Mitral Valve Prolapse

Injuries caused by free-fall from heights represent a specific form of blunt trauma that can be difficult to interpret, especially when the impact medium is water. On autopsy practice, the immersed bodies with rib fractures and internal organ injuries were often encountered and many studies have reported that impact with the water surface could cause skeletal fracture and visceral organ rupture. The height of the fall and body orientation on impact are the most important factors determining the severity and range of these injuries. In the present case, the victim was a 69-year-old male who weighed 48 kg. Following were the autopsy findings: numerous pin-point epidermal injuries on the body surface (like those due to nibbling by fish), a massive soft tissue hemorrhage in the left upper chest, multiple rib fractures (3rd–7th left ribs), ballooning of the lungs, froth in the trachea and bronchus, pericardial tearing in the right anterolateral side (7-cm long), intra-pericardial hemorrhage, and focal hemorrhage in the adventitia of the intra-pericardial aorta. No sign of an external wound was observed in the left chest area. The cause of death was drowning, and the manner of death was suicide. Unfortunately, the site and height of fall were not investigated.
Adventitia ; Aged ; Aorta ; Autopsy ; Bronchi ; Cause of Death ; Drowning ; Hemorrhage ; Humans ; Lung ; Male ; Rib Fractures ; Rupture ; Suicide ; Tears ; Thorax ; Trachea ; Water ; Wounds and Injuries

PURPOSE: Adventitial cystic disease (ACD) is a rare condition that causes intermittent claudication and non-atherosclerotic disease without cardiovascular risk factors. The etiology and optimal treatment of ACD remain controversial. The purpose of this study was to analyze surgical treatment results for ACD and to elucidate optimal treatment options.MATERIALS AND METHODS: We retrospectively reviewed 30 patients with ACD who underwent surgery from 2006 to 2018. Twenty-two patients had arterial ACD, six had venous ACD, and two had combined venous and arterial ACD. We reviewed demographic and clinical characteristics, treatment details, and procedure outcomes.RESULTS: Recurrence occurred in 6 cases either after cyst excision alone (4/17) or patch angioplasty (2/2). There was no recurrence after vessel excision with interposition grafting (0/7). Therefore, vessel excision was a statistically significant factor in recurrence prevention (P=0.026). Among the six recurrences, joint connections of the cystic lesions were found in four of the six (66.7%).CONCLUSION: As a curative surgery for ACD, vessel excision with interposition grafting is a better strategy to prevent recurrence than simple cyst excision alone.
Adventitia ; Angioplasty ; Humans ; Intermittent Claudication ; Joints ; Recurrence ; Retrospective Studies ; Risk Factors ; Transplants

More than 700 adventitial cystic diseases (ACDs) have been reported in the literature, with most cases affecting the popliteal artery in young men. Here, we describe our treatment and etiologic consideration of a patient who presented with an ACD of the external iliac artery, known to be an extremely rare location. On preoperative imaging, the ACD had a connection to the nearby hip joint and was treated with resection of the affected segment, including ligation of the joint connection and interposition with a prosthetic graft. The pathogenesis of ACDs is not fully understood; however, we believe that joint connections are important in their development and treatment.
Adventitia ; Hip Joint ; Hip ; Humans ; Iliac Artery ; Joints ; Ligation ; Male ; Popliteal Artery ; Transplants

Two patients were admitted to our department because of recent aggravation of claudication in the leg, which was exacerbated by walking. They were diagnosed as having a Baker cyst or acute thrombosis in the popliteal fossa at another hospital. There was no evidence of ischemia, and the ankle brachial index was normal. Computed tomography and magnetic resonance imaging were performed, revealing a cystic mass of the popliteal artery (PA). Intraoperatively, the cystic lesion was found within the adventitia of the PA; based on the biopsy findings, both patients were diagnosed as having adventitial cystic disease of the PA.
Adventitia ; Ankle Brachial Index ; Biopsy ; Humans ; Ischemia ; Leg ; Magnetic Resonance Imaging ; Popliteal Artery ; Popliteal Cyst ; Thrombosis ; Walking